Target:
TARGET NAME: UDP-N-acetylglucosamine pyrophosphorylase
GENE NAME: AFUA_7G02180

NCBI Gene # or RefSeq#:
70982372
66844338

Protein ID (NP or XP #) or Wolbachia#:
4BMA.10.2210

Organism:
Scientific Name: Neosartorya fumigata
Also Known As: Aspergillus fumigatus
Strains: ATCC MYA-4609 / Af293 / CBS 101355 / FGSC A1100)

Etiologic Risk Group (see link below):
Risk Group 2

Background/Disease Information:

Neosartorya fumigata is most commonly known as Aspergillus fumigatus and is the predominant cause allergic and invasive aspergillosis. The fungus is a primary pathogen of the sinuses, lungs, damaged skin, and subcutaneous tissue. It has the ability to spread to other organs including the brain. The medications used against

Aspergillus fumigatus are azoles like voriconazole, however the fungus is beginning to build resistance to the drugs. In fungus the synthasis of chitin is an essential component of the cell wall. The sugar nucleotide UDP-N-acetylglucosamine (UDP-GLCNAc) is essential in the production of this chitin. The enzymeUDP-N-acetylglucosamine pyrophosphorylase is the enzyme that is essential in the production of the sugar nucleotide (UDP-GLCNAc). In conclusion, this enzyme is a key target when it comes to the Aspergillus fumigatus disease. The general idea is to inhibit the enzyme, which will stop the production of the essential sugar nucleotide, then no chitin will be made, and the cell wall will corrupt killing the fungus.


a_fumigatus_s.jpg
Figure 1: Microscopic image of Aspergillus fumigatus


F1.large.jpg

Figure 2: How Aspergillus fumigatus invades the lungs

Link to TDR Targets page (if present):
There were no links for the organism Aspergillus fumigatus. The link given below is for other organisms.

Link to Gene Database page:
There were no links for the organism Aspergillus fumigatus on Gene Database

I was having trouble finding a link for the organism I chose. The links provided show the enzyme with other organisms. The closest I got was this:

Essentiality of this protein:
Inferred from electronic annotation. Source: UniProtKB-EC

Complex of proteins?: None were given.

Druggable Target: None were given.

EC#: 2.7.7.23

Link to BRENDA EC# page:

Screenshot of BRENDA enzyme mechanism schematic There was not one present.

Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Link to Sigma page for assay (see Sigma links below):

Product Number: U4375
CAS Number: 91183-98-1
EC Number: 293-820-5
MDL Number: MFCD00799985
Supplier: Sigma-Aldrich

Pack Size
Price (USD)
25 MG
54.60
100 MG
185.50
500 MG
564.00
1G
972.00

Structure Available (PDB or Homology model) PDB #: 4BMA

Current Inhibitors GOL- Glycerol
More info on this website:

Expression Information: Has been expresses in Escherichiu coli

Purification Method:
http://www.ncbi.nlm.nih.gov/pubmed/17142897

Image of protein:

protein discovery protein.jpg


Figure 3: PDB image of the UDP-N-acetylglucosamine diphosphorylase enzyme
Amino Acid Sequence: MAVAIKETVSNFMERFHGHPENLPREPSAEEFQQLRKKYTDAGQGHVFAFV
DELQTGERSQLFHQLSSFDPVRINELADKALNPPKADDGPASLEPLPDITASILDSDPKDLEQWYEEGLKLVAGNKVAVVLMAGGQGTRLGSSAPKGCFDIGLPSHKSLFQIQAERIAKLQLLAQRISG.KEAVIPWYVMTSGPTRKPTEEFFEQHKYFGLNKSDVIIFEQGVLPCISN.EGKILMESKFKVAVAPDGNGGIYQALLTSGVREDMRKRGIEHIHTYCVD.NCLVKVADPVFIGFAASKQVDIATKVVRKRNATESVGLILQKNGKPDVVE.YSEIDKETAEAKDPKQPDVLKFRAANIVNHYYSFKFFESIELWAHKLPHH.VARKKIPCIKEGTGEFFKPEKPNGIKLEQFVFDVFPMTPLEKFACIEVRR.EDEFSPLKNARGTGEDDPDTSKRDIMSQGQRWIEKAGGIVITEGDVVGVEVSPLISYGGEGLEFLKGREIKAPAFIEKEE

Length of your Protein in Amino Acids:
509

Molecular Weight:
56715.7kDal

Molar Extinction coefficient of your protein at 280 nm wavelength Assuming all pairs of Cys residues form cystines: 37275
Assuming all Cys residues are reduces: 3690

TMpred Graph Image:Graph for protein.gif


CDS Gene Sequence (paste as text only):


GC% Content for gene:
Not given.

CDS Gene Sequence (codon optimized) - copy from output of Primer
Primer did not generate the information.

Design Protocol (paste as text only):
Primer did not generate the information.

GC% Content for gene (codon optimized):
Primer did not generate the information.

Resources:

See ProtocolTargetDiscoveryVDS.docx for more
Etiologic Risk Group Categories (for pathogens):http://www.utexas.edu/research/rsc/ibc/agent_class.html#_Toc7238334

SIGMA-ALDRICH RESOURCES
Enzyme Explorer
http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer.html

Enzyme Classification Index (EC number)
http://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=14573088


WolframAlpha http://www.wolframalpha.com/
DrugBank http://www.drugbank.ca/


Databases of genes/organisms:
http://www.niaid.nih.gov/Pages/default.aspx
http://eupathdb.org/eupathdb/
https://patricbrc.vbi.vt.edu/portal/portal/patric/Home
http://www.nmpdr.org/FIG/wiki/view.cgi/Main/EssentialGenes
http://tubic.tju.edu.cn/deg/
http://csgid.org/csgid/cake/pages/community_request_gateway
http://tdrtargets.org/
http://gsc.jcvi.org/status.shtml


Scientific Nomenclature page from Center for Disease Control (gene, protein names and abbreviations)
http://wwwnc.cdc.gov/eid/pages/scientific-nomenclature.htm


Gene Information:
NCBI GENE Page: http://www.ncbi.nlm.nih.gov/gene
BLAST Page: http://blast.ncbi.nlm.nih.gov/

Protein Information:
NCBI Protein Page: http://www.ncbi.nlm.nih.gov/protein
Protein Expression Website
Protein Expression Paper: SGC_ProteinProductionPurificationNatMethods2008.pdf

Primer Overlap PCR Articles
HooverLubkowski_PCRoverlapcloninggnf042.pdf
StemmerPCRoverlapGene1995.pdf

Is my target good for Virtual Screening programs?
Reynolds_THermodynamicsLigandBinding_MedChemLett2011.pdf